Modulating the Alternative Complement Pathway

a technology of alternative complement and pathway, applied in the field of modulating the alternative complement pathway, can solve the problems of insufficient local control of the alternative pathway by the target tissue, and the limited ability of circulating factor h to protect the surface of hypoxic tubular epithelial cells

Active Publication Date: 2011-09-22
UNIV OF COLORADO THE REGENTS OF
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0008]This application pertains to methods and compositions for modulating, e.g., stimulating or inhibiting, activity of the alternative complement pathway.

Problems solved by technology

Injury to host tissues by the alternative pathway indicates insufficient local control of the alternative pathway by the target tissue.
Nevertheless, uncontrolled activation of the alternative pathway in the setting of reduced surface Crry indicates that circulating factor H has a limited ability to protect the surface of hypoxic tubular epithelial cells.

Method used

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  • Modulating the Alternative Complement Pathway
  • Modulating the Alternative Complement Pathway
  • Modulating the Alternative Complement Pathway

Examples

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example 1

Factor H Purification, Analysis and Activities

Materials and Methods

[0225]Purification of mouse factor H. An affinity column for factor H was made by ligating polyclonal goat antibody for human factor H (Quidel Corp., San Diego, Calif.) to Sepharose® derivatized with cyanogen bromide (CNBr) (Amersham Biosciences / GE Healthcare, Piscataway, N.J.) according to the manufacturer's instructions. Plasma was collected from C57BL / 6 mice and passed over the column. After washing the column with phosphate buffered saline (PBS), pH 7.4, the factor H was eluted with 5 M LiCl2. The salt solution was exchanged with PBS and the factor H was then passed through a Sephadex™ 26 / 60 Superdex™ 200 column (Amersham Biosciences / GE Healthcare, Piscataway, N.J.). The purity and identity of the isolated protein was verified by Coomassie staining and Western blot analysis by standard methods.

[0226]Western blot and Far Western blot analysis. Renal tissue was homogenized in radioimmunoprecipitation assay (RIPA) l...

example 2

Effect of Annexin 2 During Renal Failure

Experimental

[0253]Renal ischemia / reperfusion protocol. Eight to ten week old male C57BL / 6J mice (Jackson Laboratories) mice weighing 20-25 g were anesthetized with 60 mg / kg ketamine plus 10 mg / kg xylazine (Vedco, Inc., St. Joseph, Mo.) injected intra-peritoneally. Mice were placed on a heating pad to maintain body temperature during surgery. Laparotomies were then performed and the renal pedicles were located and isolated by blunt dissection as known in the art. The pedicles were clamped with surgical clips (Miltex Instrument Company), and occlusion of blood flow was confirmed by visual inspection of the kidneys. The clamps were left in place for 24 minutes and then released. The kidneys were observed for approximately one minute to ensure blood re-flow, then fascia and skin were sutured with 4-0 silk (United States Surgical). Sham surgery was performed in an identical fashion, except that the renal pedicles were not clamped. The mice were vol...

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Abstract

Provided herein are compositions, including pharmaceutical compositions, and methods for modulating, i.e., stimulating or inhibiting, activity of the alternative complement pathway, and methods of identifying factor H-binding proteins.

Description

CROSS-REFERENCES TO RELATED APPLICATIONS[0001]This is a national stage of International Application No. PCT / US2009 / 057912, filed Sep. 22, 2009, which claims the benefit of U.S. Provisional Application No. 61 / 099,173, filed Sep. 22, 2008, the entire contents of which are incorporated herein by reference in their entireties and for all purposes.STATEMENT OF RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT[0002]This invention was made with government support under grant numbers K08 DK064790, R01 AI031005 and R01 DK076690 awarded by the National Institutes of Health. The government has certain rights in the invention.BACKGROUND OF THE INVENTION[0003]The alternative pathway of complement is a phylogenetically ancient arm of the innate immune system that eliminates invasive pathogens and facilitates the removal of injured host cells. See J. M. Thurman et al., J. Immunol. (2006) 176:1305-1310. The alternative pathway is continually auto-activated in the fluid pha...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K39/395A61P43/00
CPCA61K38/1709A61K38/00C07K14/4721A61K38/177C12N9/88C12Y402/01002A61P43/00C07K16/18C07K2317/54C07K2317/55C07K2319/70
Inventor THURMAN, JOSHUA M.HOLERS, V. MICHAEL
Owner UNIV OF COLORADO THE REGENTS OF
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